Toxic Pseudo-nitzschia multistriata (Bacillariophyceae) from the Gulf of Naples: morphology,toxin analysis and phylogenetic relationships with other Pseudo-nitzschia species

The genus Pseudo-nitzschia includes several species capable of producing domoic acid, the causative agent of Amnesic Shellfish Poisoning. Some of these species have been recorded frequently in the Gulf of Naples. For one of the species, P. multistriata, which has been recurrently found in our sampling area since 1995, this is the first report for European waters. Here we provide further details on the fine structure of this species. Pseudo-nitzschia multistriata was the only one found to produce domoic acid among all the Pseudo-nitzschia species from the Gulf of Naples, and this finding raises the number of potentially toxic species in this genus to nine. Phylogenetic relationships among several Pseudo-nitzschia species were assessed using the hypervariable domains (D1–D3) of the large subunit (LSU) rDNA. The match between the phylogeny obtained and important taxonomic characters used in this genus are discussed. Results show that P. multistriata clusters with wider species lacking a central larger interspace in the raphe. Close genetic relationships were determined between P. fraudulenta and P. subfraudulenta, and between P. pungens and P. multiseries. Genetic differences among these pairs of species are comparable to those among isolates of P. pseudodelicatissima from the Gulf of Naples, indicating high intraspecific genetic diversity of Pseudo-nitzschia species in the relatively conserved LSU region. This could explain the problematic results obtained when testing a match between species-specific Pseudo-nitzschia LSU probes and our sequences.     

Evaluation of cadmium,lead and metallothionein contents in the tissues of mussels (Mytilus galloprovincialis) from the Campania coast (Italy): Levels and seasonal trends

The biological effect of seasonality on cadmium, lead and metallothionein contents was assessed in mussels Mytilus galloprovincialis from natural banks located along the coastline of the Gulf of Naples (Campania, Italy). Heavy metals and metallothionein concentrations were measured in digestive and reproductive glands. The results showed a clear correlation between metallothionein content and the reproductive gland status determined during the seasons; on the contrary, no correlation was found between metallothionein and metal contents. Data allow us to hypothesize that metallothionein functions go beyond metal detoxification, thus opening new scenarios for these proteins in invertebrates. The effect of seasons on metals concentration in mussel tissues showed similar seasonal patterns between the sites, regardless of their anthropogenic impacts. Cadmium content was not strictly related to seasonal periods, whereas lead content was significantly lower in summer. The results also indicate that the metal contents in mussels from the Gulf of Naples do not represent a risk to human health, even in the period of their maximum accumulation, and that the relaying of mussels before marketing could improve the animal stress conditions, but having a slight effect on metal excretion.     

Phylogeography of the sea urchin Paracentrotus lividus (Lamarck) (Echinodermata:Echinoidea): first insights from the South Tyrrhenian Sea

The sea urchin Paracentrotus lividus (Lamarck) (Echinodermata: Echinoidea) is an Atlanto-Mediterranean species abundant in the littoral zone, where it occurs in the sublittoral down to 20 m. The aim of our work is to investigate the genetic patterns of P. lividus along the South Tyrrhenian coasts. Five specimens were collected in six localities, from the Gulf of Naples and the Cilento coast. The nuclear rDNA ITS2 spacer and the two mitochondrial genes 16S and COI were used for the analysis. The three markers utilised did not show any structure among populations from the Gulf of Naples. All populations appear to be polyphyletic, with Cilento samples more differentiated from the others. This suggests the existence of phylogeographic structure at larger geographic scale. Absence of genetic structure has been interpreted taking into consideration theoretical dispersal of the planktotrophic larvae, which can survive for 4–8 weeks before settlement, marine current patterns and persistence of the species in the area.     

Structural analysis of winter phytoplankton in the Gulf of Naples

Quantitative and qualitative distribution of surface phytoplankton,as related to hydrographic conditions, was studied in the Gulfof Naples in February 1979. Previous work has shown that the Gulf of Naples is a diversifiedecosystem, due to geographic and hydrographic features as wellas man made eutrophication, that can be subdivided into twomajor parts: a coastal subsystem and an open water one. Hydrographic analysis of the winter situation at the surfacefully confirms this picture, as it identifies two distinct watermasses corresponding respectively to surfaced Tyrrhenian IntermediateWater and to Coastal Surface Water. The structural analysisof phytoplankton reveals three assemblages of species characterizingdifferent water types: 1 - the Ischia and Procida channels affectedby the advection of Volturno river and Cuma outfall plumes;2 - the coastal area of the Gulf proper, namely the bays ofPozzuoli, Naples and Castellammare; 3 - the open waterhemislocated beyond the 100 m isobath. The channel area assemblage is dominated by diatoms, particularlyby fast growing species, such as Asterionella japonica, severalspecies of Chaetoceros and Rhizosolenia hebetata f. semispina.The coastal assemblage is identified, among others, by the diatomsCerataulina bergonii, Hemiaulus sinensis; the dinoflagellatesGlenodinium lenticula, Exuviaella compressa and Porella perforata.The open water assemblage is characterized by the diatoms Coscinodiscuscurvatulus and Hemidiscus cuneiformis, the dinoflagellate Amphidiniumacutissimum and the coccolithophore Coccolithus haeckelii.     

Teredoika aspectabilis sp. n., a new endoparasitic copepod from the gastropod mollusc, Clinopegma unicum, in the Sea of Okhotsk

A new species of the genus Teredoika Stock, 1959 is described. T. aspectabilis sp. n., the second species of the genus, was found in the kidney of Clinopegma unicum in the Okhotsk Sea while the type species is known from the stomach of the shipworm Teredo utriculus caught in the Gulf of Naples.     

On the identity of a Centaurea population on Procida Island,Italy: Centaurea corensis rediscovered

The discovery on the Island Procida (Gulf of Naples, off the Italian west coast) of a Centaurea population similar to C. corensis, a narrow-range Sardinian endemic hitherto known only from one single locality in Sardinia, is presented here. The identification of the two populations as C. corensis is confirmed by morphological comparison and morphometric analysis, chromosome counts and nuclear ribosomal DNA sequence data. A recent origin of the species through polyploidization is hypothesized. Finally, the possibility that recent anthropogenic dispersal may account for the disjunction is discussed.     

Temporal Changes in Population Structure of a Marine Planktonic Diatom

A prevailing question in phytoplankton research addresses changes of genetic diversity in the face of huge population sizes and apparently unlimited dispersal capabilities. We investigated population genetic structure of the pennate planktonic marine diatom Pseudo-nitzschia multistriata at the LTER station MareChiara in the Gulf of Naples (Italy) over four consecutive years and explored possible changes over seasons and from year to year. A total of 525 strains were genotyped using seven microsatellite markers, for a genotypic diversity of 75.05%, comparable to that found in other Pseudo-nitzschia species. Evidence from Bayesian clustering analysis (BA) identified two genetically distinct clusters, here interpreted as populations, and several strains that could not be assigned with ≥90% probability to either population, here interpreted as putative hybrids. Principal Component Analysis (PCA) recovered these two clusters in distinct clouds with most of the putative hybrids located in-between. Relative proportions of the two populations and the putative hybrids remained similar within years, but changed radically between 2008 and 2009 and between 2010 and 2011, when the 2008-population apparently became the dominant one again. Strains from the two populations are inter-fertile, and so is their offspring. Inclusion of genotypes of parental strains and their offspring shows that the majority of the latter could not be assigned to any of the two parental populations. Therefore, field strains classified by BA as the putative hybrids could be biological hybrids. We hypothesize that P. multistriata population dynamics in the Gulf of Naples follows a meta-population-like model, including establishment of populations by cell inocula at the beginning of each growth season and remixing and dispersal governed by moving and mildly turbulent water masses.     

A NEW MEDITERRANEAN SPECIES OF HAMINAEA LEACH, 1820 (GASTROPODA: OPISTHOBRANCHIA:CEPHALASPIDEA)

A new species of cephalaspidean gastropod from MediterraneanSea (West Naples Gulf, Italy) is described. This species, Haminaeafusari, is characterised mainly by the presence of short parapodiallobes, which cover only the anterolateral parts of the shell;the prostatic gland has two lobes separated by a narrow andcylindrical zone; the lateral teeth of the radula are all smooth;the penis has a triangular section at the apical zone and asmall sharpened tip; and the spawn is horse-shoe-shaped or coiledin a single spiral. This species is compared with all the Europeanspecies of the genus and a key to identification is provided. (Received 4 May 1992; accepted 4 January 1993)     

Nocardiosis in Mediterranean bivalves: First detection of Nocardia crassostreae in a new host Mytilus galloprovincialis and in Ostrea edulis from the Gulf of Naples (Italy)

In this work M. galloprovincialis and O. edulis specimens were surveyed for a pathological study in the Gulf of Naples (Mediterranean sea, Campania Region, southern Italy). Clusters of Nocardia sp.-like cells were observed in histological slides. PCR amplification, sequencing and in situ hybridization were carried out in order to corroborate Nocardia species identification for both hosts. Blast results showed a 99% of maximum identity with Nocardia crassostreae sequences in Genbank. This is the first report of N. crassostreae in the new host M. galloprovincialis and, in a new area, the Mediterranean Sea.     

Mass mortality in Northwestern Mediterranean rocky benthic communities: effects of the 2003 heat wave

Late in summer 2003, extensive mass mortality of at least 25 rocky benthic macro-invertebrate species (mainly gorgonians and sponges) was observed in the entire Northwestern (NW) Mediterranean region, affecting several thousand kilometers of coastline. We were able to characterize the mortality event by studying six areas covering the main regions of the NW Mediterranean basin. The degree of impact on each study area was quantified at 49 sites by estimating the proportion of colonies affected in populations of several gorgonian species compared with reference data obtained in years without mortality signs. According to these data, the western areas (Catalan coast and Balearic Islands) were the least affected, while the central areas (Provence coast and Corsica-Sardinia) showed a moderate impact. The northernmost and eastern areas (Gulf of Genoa and Gulf of Naples) displayed the highest impact, with almost 80% of gorgonian colonies affected. The heat wave of 2003 in Europe caused an anomalous warming of seawater, which reached the highest temperatures ever recorded in the studied regions, between 1 and 3 °C above the climatic values (mean and maximum). Because this exceptional warming was observed in the depth ranges most affected by the mortality, it seems likely that the 2003 anomalous temperature played a key role in the observed mortality event. A correlation analysis between temperature conditions and degree of impact seems to support this hypothesis. Under the present climate warming trend, new mass mortality events may occur in the near future, possibly driving a major biodiversity crisis in the Mediterranean Sea.     

Histomorphological changes in the reproductive condition of parasitized marine planktonic copepods

Changes in the internal morphology of parasitized copepods sampledin the Gulf of Naples from 1986 to 1988 were investigated usingboth light and transmission electron microscopy. The most commonand devastating form of infection was due to the parasitic dinoflagellateSyndinium, which induced gross modifications in the internaland external morphology of the host population. The parasitereduced fecundity and longevity of infected individuals, whichmainly included juvenile and female populations of the copepodParacalanus parvus. Infection rates for this species were ashigh as 12% for. juveniles and 13% for adult females for theperiod investigated. Infection by the parasitic dinoflagellateBlastodinium induced less negative effects on the reproductivebiology of their hosts. This pathogen did not lead to sexualcastration and oogonal development appeared normal. Other formsof infestation were rare and included infections due to protozoa,fungi and bacteria. ^*Died tragically at sea during a cruise on December 15, 1988     

Red cell glyoxalase I polymorphism in Italians. Report of a variant phenotype.

The glyoxalase I polymorphism was studied in 1,490 unrelated subjects from three different areas of Italy (Milan, Rome and Naples). The following frequencies of GLO2 allele were observed: Milan 0.57, Rome 0.59, Naples 0.61, and a significant difference between Milan and Naples was found (p less than 0.02). An anomalous phenotype is also described which permits us to postulate a new allele producing an isozyme with low activity and electrophoretically slower than those corresponding to GLO1 and GLO2 alleles.     

Seasonal fluctuations in fecundity and hatching success in the planktonic copepod Centropages typicus

A mismatch of {small tilde}2–3 months was observed betweenreproductive rates and population abundances in the planktoniccopepod Centropages typicus, with the highest production atrelatively low female abundance and low production at high abundance,during the course of a 2 year study in 1989 and 1990 in neriticwaters of the Gulf of Naples. During this period, egg mortalitywas at times severe, with values as high as 59% in February1989. The seasonal trend in percentage hatching success didnot match seasonal fluctuations in breeding intensity, and wasnot correlated with variations in environmental variables suchas temperature and chlorophyll a. The results of experimentsusing the fluorescent dye Hoechst 33342 showed that unhatchedeggs had been fertilized and that in most cases developmenthad proceeded to an advanced stage before death of the embryo.The causes of high egg mortality are uncertain, but here wegive evidence, for the first time, that egg mortality may notonly be due to infertility caused by failure to remate.     

Using citizen science to obtain data on large,floating gelatinous spheres from NE Atlantic,attributed to egg mass of ommastrephid squid (Oegopsida,Cephalopoda, Mollusca)

A total of 27 large, gelatinous spherical masses observed in coastal Norwegian waters from Nordland to Aust-Agder Counties in Norway, and off Lysekil in Sweden, Muljica Island in Croatia, Gulf of Naples in Italy, Reqqa Point in Malta, and Saint Mandrier in France, during the months of April to September 2001 to 2017, are reported. Individual spheres measured 0.3–2 m in diameter, averaging one metre (n?=?24, ±0.53 m), with all but four sighted in suspension in the water column between 0.5 and 52 m depth, in water temperatures ranging between 10–21°C. About half of all spheres contained a yellow-red streak through their gelatinous core. Tissue samples were not obtained. We attribute these gelatinous spheres to the egg masses of squid (Cephalopoda, Oegopsida), and most likely to the ommastrephid Todarodes sagittatus, given similarities with egg masses of T. pacificus.     

ALEXANDRIUM TAMUTUM SP. NOV. (DINOPHYCEAE): A NEW NONTOXIC SPECIES IN THE GENUS ALEXANDRIUM1

A new species of the dinoflagellate genus Alexandrium, A. tamutum sp. nov., is described based on the results of morphological and phylogenetic studies carried out on strains isolated from two sites in the Mediterranean Sea: the Gulf of Trieste (northern Adriatic Sea) and the Gulf of Naples (central Tyrrhenian Sea). Vegetative cells were examined in LM and SEM, and resting cysts were obtained by crossing strains of opposite mating type. Alexandrium tamutum is a small‐sized species, resembling A. minutum in its small size, the rounded‐elliptical shape and the morphology of its cyst. The main diagnostic character of the new species is a relatively wide and large sixth precingular plate (6″), whereas that of A. minutum is much narrower and smaller. Contrary to A. minutum, A. tamutum strains did not produce paralytic shellfish poisoning toxins. Phylogenies inferred from the nuclear small subunit rDNA and the D1/D2 domains of the large subunit nuclear rDNA of five strains of A. tamutum and numerous strains of other Alexandrium species showed that A. tamutum strains clustered in a well‐supported clade, distinct from A. minutum.     

Potentially toxic and harmful microalgae from coastal waters of the Campania region (Tyrrhenian Sea, Mediterranean Sea)

A total number of 40 potentially toxic microalgae and 5 taxa causing discolorations have been identified along the coasts of the Campania region (South Tyrrhenian Sea, Mediterranean Sea). This number is based on results of over 20 years of research at a coastal station in the Gulf of Naples, 4 years of monitoring activity along the Campania coasts and of a series of scattered and sporadic observations. Several species of the recently erected genera Karenia and Takayama are reported for the first time in the area. Information on the period of highest abundances or most probable period of occurrence indicates the late spring and summer as the periods of maximum risk of harmful events. Despite the variety of potentially toxic species, no human health problems nor fish kills have ever been recorded in Campania. The reasons for this apparent paradox are probably to be found in the ecological factors that regulate the abundance, toxicity and spatial distribution of the potentially harmful species and reflect as well the relatively low number of aquaculture plants in the area.     

Redescription of Cardiosporidium cionae (Van Gaver and Stephan, 1907) (Apicomplexa: Piroplasmida), a plasmodial parasite of ascidian haemocytes

Cardiosporidium cionae (Apicomplexa), from the ascidian Ciona intestinalis L., is redescribed with novel ultrastructural, phylogenetic and prevalence data. Ultrastructural analysis of specimens of C. intestinalis collected from the Gulf of Naples showed sporonts and plasmodia of C. cionae within the host pericardial body. Several merogonic stages and free merozoites were found in the pericardial body, together with sexual stages. All stages showed typical apicomplexan cell organelles, i.e. apicoplasts, rhoptries and subpellicular microtubules. Merogonic stages of C. cionae were also produced inside haemocytes. A fragment of the rSSU gene of C. cionae was amplified by PCR using DNA extracted from the pericardial bodies. The amplified product showed closest affinity with other apicomplexan representatives and a 66bp unique insertion, specific for C. cionae, at position 1644. Neighbour-joining phylogenetic analysis placed C. cionae in a clade with other piroplasm genera, including Cytauxzoon, Babesia and Theileria spp. The parasite was found in different populations of C. intestinalis with highest prevalence in October-November. Ultrastructural and DNA data showed that the organism, described in 1907 from the same host but not illustrated in detail, is a member of a novel marine apicomplexan radiation of tunicate parasites.     

Metabolic fingerprinting reveals differences between northern and southern strains of the cryptic diatom Chaetoceros socialis

Morphology and molecular phylogeny constitute the structural elements of diatom taxonomy. These approaches do not, however, give information on the functioning of taxa. Additional methods to serve a more integrated and wide-ranging taxonomy have therefore been called for. Metabolic fingerprinting is one approach used within the field of metabolomics, often applied in classification of samples. Here we apply metabolic fingerprinting in a taxonomic study of a cryptic diatom species. Strains of the cosmopolitan diatom Chaetoceros socialis from two geographical areas; the north-east Atlantic and Arctic and the Gulf of Naples, were cultivated at three different temperatures; 2.5, 8 and 13°C. The strains from the two different geographical areas exhibited different growth rates as well as different photosynthetic efficiencies. Algal extracts, collected at the end of the growth experiments, were analysed by Ultra-Performance Liquid Chromatography High Resolution Mass Spectrometry. The two groups of strains were separated by principal component analysis of their metabolic fingerprints. Analysis of the data revealed both qualitative and quantitative differences in metabolite markers. These phenotypic differences reinforce differences also found for morphology, phylogenetic markers and growth rates, and point at different adaptive characteristics in organisms living under different temperature regimes.